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Immunoassays | Metabolism

Erenna® Mouse (Plate-based) Insulin Immunoassay Kit (Cat# 03-0006-xx)

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The Erenna® Insulin Mouse Immunoassay Kit uses a quantitative fluorescent sandwich immunoassay technique to measure insulin in mouse heparin plasma samples. A capture antibody specific for mouse insulin has been pre-coated onto a 384-well microplate. The user pipettes standards and samples into microplate wells along with a fluorescent-labeled dye detection antibody. During incubation, the insulin present in the sample binds to the solid phase capture antibody. Unbound insulin molecules are washed away during the subsequent wash step. This detection antibody will recognize and bind to insulin that has been captured onto the solid phase. After washing, elution buffer is added and incubated to dissociate the bound protein sandwiches from the plate surface. The fluorescent antibodies are now free-floating in the wells. The plate is loaded into the Erenna System and the fluorescent molecules are counted. The number of fluorescently labeled detection antibodies counted is directly proportional to the amount of insulin that was present in the sample when captured. The amount of insulin in unknown samples is interpolated off of the standard curve.

Assay Performance

TABLE 1. Analytical sensitivity of the Erenna® Mouse (Plate-based) Insulin Immunoassay Kit1
Lower Limit of Detection 12 pg/mL
Lower Limit of Quantification2 20 pg/mL
Upper Limit of Quantification 1280 pg/mL
Low-end CV% Range 3 - 16%
Low-end CV% Average 13%
Recommended Sample Volume 5 μL

1 See product insert for updated values
2 LLoQ = 20% CV and ± 20% recovery

FIGURE 1. Unlike two other commercial ELISAs, the Erenna® Mouse, plate-based Insulin Immunoassay Kit shows good precision and recovery. § ELISA 1: HRP plate-based, 5 μL assay volume † ELISA 2: HRP plate-based, 10 μL assay volume

FIGURE 2. Assay linearity among the Erenna® Mouse, plate-based Insulin Immunoassay Kit and two other ELISAs (left), and a close up of the Erenna® Mouse, plate-based Insulin Immunoassay Kit low-end standard curve (right). The Erenna® Mouse, plate-based Insulin Immunoassay Kit offers superior linearity, robust precision (CV ≤ 20%) and excellent recovery (± 20%) as opposed to other commercially available ELISAs, allowing for accurate measurement of insulin in mouse.

Representative data shown for demonstration purposes only. Individual results may vary depending upon samples tested and protocol used.

Biology and Disease

Insulin is an important metabolic hormone produced by islet cells within the pancreas whose primary function is to regulate blood sugar levels in the body. When blood glucose level is high (ie. after food ingestion), insulin increases glucose uptake and forces liver cells to convert glucose to glycogen for storage. Insulin is also known to promote glycolysis, fatty acid synthesis and amino acid uptake. Patients who do not produce sufficient insulin (Type 1) or become resistant to insulin (Type 2) develop diabetes mellitus and suffer from hyperglycemia, which can lead to complications such as blindness, cardiovascular disease, and kidney damage. Many insulin analogs, which mimic either normal or rapid-acting (ie. lispro) insulin functions have been developed as treatment for diabetes.

TABLE 2. Additional UniProtKB/Swiss-Prot Information*
Protein Name: Insulin-1
UniProtKB/Swiss-Prot ID: P01325
Protein-protein interaction database: P01325
Alternative Names:
Gene Names: Ins1
Synonym Gene Names: Ins-1
Function: Insulin decreases blood glucose concentration. It increases cell permeability to monosaccharides, amino acids and fatty acids. It accelerates glycolysis, the pentose phosphate cycle, and glycogen synthesis in liver.
Biological Processes: carbohydrate metabolism, glucose metabolism
Molecular Function: hormone activity

* Source Info from www.uniprot.org.

Related Products

Analyte Description  Catalog #
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GLP-1, total (MP-based) Erenna® Total GLP-1(MP-based) Immunoassay Kit  03-0025-xx
GLP-1, total (Plate-based) Erenna® Total GLP-1(Plate-based) Immunoassay Kit  03-0066-xx